人外周血自然杀伤T细胞分泌胰岛素样生长因子-1的研究

Expanding NKT Cells in Vitro and Detecting the Release of IGF-1 from the Cells

目的：建立人外周血自然杀伤T细胞（NKT）体外扩增及检测NKT细胞分泌胰岛素样生长因子-1（IGF-1）的方法。方法：采用IFN-γ，CD3和IL-2的方法从人外周血单个核细胞（PBMC）中扩增出NKT细胞，采用流式细胞仪检测扩增效率，并使用绝对实时定量PCR（real-timePCR）的方法检测扩增出的NKT细胞中IGF-1的mRNA表达量。使用内源性管家基因GAPDH校正误差。结果：两周扩增后NKT细胞增加了4倍。受试者扩增的NKT细胞IGF-1mRNA表达量为1．29-I-0．56拷贝数／10^6GAPDH。结论：从PBMC扩增NKT细胞可以获得良好的效果，扩增出的NKT细胞有IGF-1的表达，但是表达量相对较低。

Objective To expand the NKT cells in vitro and to determine the release of IGF- 1 from the NKT cells. Methods NKT cells were expanded from human peripheral blood mononuclear cell （PBMC）. Flow cytometry was used to determine the expanding efficiency. Gene expression of IGF - 1 mRNA in expanding NKT cells was detected by real - time PCR. An endogenous ＇ housekeeping＇ gene GAPDH was used to normalize the results. Results After two - week expansion, 4 - fold increase in the number of NKT cells was found. Gene expression of IGF - 1 mRNA in expanding NKT cells was 1.29 ＋ 0,56 copies/10^6 GAPDH. Conclusions The NKT cells can be expanded very well in vitro from PBMC and NKT cells can express IGF - 1 as well, but the expression is comparatively low.